Kobayashi T, Kato Y, Tsunoda Y
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan.
Theriogenology. 2004 Sep 1;62(5):854-60. doi: 10.1016/j.theriogenology.2003.12.032.
The present study examined whether the timing of the first cleavage has an effect on the in vitro and in vivo developmental potential of nuclear-transferred mouse oocytes receiving embryonic stem cells. First, the timing of the first cleavage and the developmental potential of nuclear-transferred oocytes were examined every hour from 12 to 24 h after the start of culture and compared with in vitro-fertilized oocytes. The developmental potential of in vitro-fertilized oocytes decreased gradually according to the time required for cleavage (84% (32/38) for 15 h to 50% (1/2) for 20 h), but intermediate-cleaved (15-16 h) nuclear-transferred oocytes had a higher potential to develop into blastocysts (55% (17/31) to 67% (45/67) versus 0-43% (6/14)]. Second the nuclear-transferred oocytes were divided into three groups according to the timing of the first cleavage; each group was cultured to blastocysts in vitro, and then transferred to recipients. The potential of intermediate-cleaved oocytes (15-16 h) to develop into blastocysts was significantly higher than fast-cleaved (before 15 h) and slow-cleaved (after 16 h) oocytes (65, 46, and 37%). The proportion of fetuses on Day 10.5 of pregnancy was highest in the intermediate-cleaved group (4 versus 2 and 1%, respectively) and a full-term fetus was obtained from this group. The present study demonstrated that the timing of the first cleavage could be used to determine the potential of nuclear-transferred oocytes with embryonic stem cells to develop to the blastocyst stage in vitro, but not to determine post-implantation viability after transfer to recipients.
本研究探讨了首次卵裂时间是否会影响接受胚胎干细胞的核移植小鼠卵母细胞的体外和体内发育潜能。首先,在培养开始后的12至24小时内,每小时检查一次首次卵裂时间和核移植卵母细胞的发育潜能,并与体外受精的卵母细胞进行比较。体外受精卵母细胞的发育潜能随着卵裂所需时间逐渐降低(15小时时为84%(32/38),20小时时为50%(1/2)),但中等卵裂(15 - 16小时)的核移植卵母细胞发育成囊胚的潜能更高(55%(17/31)至67%(45/67),而对照为0 - 43%(6/14))。其次,根据首次卵裂时间将核移植卵母细胞分为三组;每组在体外培养至囊胚,然后移植到受体体内。中等卵裂(15 - 16小时)的卵母细胞发育成囊胚的潜能显著高于快速卵裂(15小时之前)和缓慢卵裂(16小时之后)的卵母细胞(分别为65%、46%和37%)。妊娠第10.5天时,中等卵裂组的胎儿比例最高(分别为4%,而其他两组为2%和1%),并且从该组获得了一只足月胎儿。本研究表明,首次卵裂时间可用于确定带有胚胎干细胞的核移植卵母细胞在体外发育至囊胚阶段的潜能,但不能用于确定移植到受体后植入后的存活能力。